Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan.
Plant Cell. 2010 Sep;22(9):3105-17. doi: 10.1105/tpc.109.071811. Epub 2010 Sep 24.
Aquatic photosynthetic organisms can modulate their photosynthesis to acclimate to CO₂-limiting stress by inducing a carbon-concentrating mechanism (CCM) that includes carbonic anhydrases and inorganic carbon (Ci) transporters. However, to date, Ci-specific transporters have not been well characterized in eukaryotic algae. Previously, a Chlamydomonas reinhardtii mutant (lcr1) was identified that was missing a Myb transcription factor. This mutant had reduced light-dependent CO₂ gas exchange (LCE) activity when grown under CO₂-limiting conditions and did not induce the CAH1 gene encoding a periplasmic carbonic anhydrase, as well as two as yet uncharacterized genes, LCI1 and LCI6. In this study, LCI1 was placed under the control of the nitrate reductase promoter, allowing for the induction of LCI1 expression by nitrate in the absence of other CCM components. When the expression of LCI1 was induced in the lcr1 mutant under CO₂-enriched conditions, the cells showed an increase in LCE activity, internal Ci accumulation, and photosynthetic affinity for Ci. From experiments using indirect immunofluorescence, LCI1-green fluorescent protein fusions, and cell fractionation procedures, it appears that LCI1 is mainly localized to the plasma membrane. These results provide strong evidence that LCI1 may contribute to the CCM as a component of the Ci transport machinery in the plasma membrane.
水生光合生物可以通过诱导碳酸酐酶和无机碳 (Ci) 转运体来调节光合作用,以适应 CO₂限制应激。然而,迄今为止,真核藻类中的 Ci 特异性转运体尚未得到很好的描述。先前,已经鉴定出一种缺失 Myb 转录因子的莱茵衣藻突变体 (lcr1)。当在 CO₂限制条件下生长时,该突变体的光依赖性 CO₂气体交换 (LCE) 活性降低,并且不会诱导编码质膜碳酸酐酶的 CAH1 基因以及两个尚未被描述的基因 LCI1 和 LCI6 的表达。在这项研究中,LCI1 被置于硝酸还原酶启动子的控制之下,允许在没有其他 CCM 成分的情况下通过硝酸盐诱导 LCI1 的表达。当在富含 CO₂的条件下诱导 lcr1 突变体中的 LCI1 表达时,细胞表现出 LCE 活性、内部 Ci 积累和对 Ci 的光合亲和力增加。通过间接免疫荧光、LCI1-绿色荧光蛋白融合和细胞分级分离程序的实验表明,LCI1 主要定位于质膜。这些结果提供了有力的证据,表明 LCI1 可能作为质膜 Ci 转运机制的一部分,作为 CCM 的组成部分。